Predicted band size: 90 kDaObserved band size: 90 kDaAdditional bands at: 38 kDa. We are unsure as to the identity of these extra bands.

Exposure time: 20 minutes

Immunoprecipitation - Anti-RRM1 antibody (ab81085)

RRM1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to RRM1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab81085.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697)..Band: Band: 90kDa: RRM1.